FIG. 6.

EB^KDM5b retain highly proliferative cells that are refractory to differentiation. (A) Cells from dissociated normal EBs after four passages. (B) Cells from dissociated EB^KDM5b after two passages. (C) Cells from dissociated normal EBs after two passages stained with DAPI (blue), Oct4 (green), and Sox1 (red). *, shows background spots. (D) Cells from dissociated EB^KDM5b after four passages stained with DAPI (blue), Oct4 (green), and Sox1 (red). *, shows background spots (C and D). Cells from dissociated EB^KDM5b after four passages stained with DAPI (blue), Nestin (green), and Nanog (red). (E) Arrow shows Nanog, Nestin double-negative colony of cells. (F) Neural differentiation of day 9 EBs from control; the edge of the colony is marked by a dashed line. Arrows show Sox1-positive (red) cells and DAPI-stained (blue) cells. (G) Neural differentiation of day 9 EBs^KDM5b; the edge of the colony is marked by dashed lines. Dashed arrow shows secondary colony forming. Arrows show Sox1-positive (red) cells and DAPI-stained (blue) cells. (H) Graph of 7-amino-actinomycin D staining of cells found in media during neural differentiation of day 9 EBs from control (normal mESCs) and mESC^KDM5b. The experiment was done in triplicate.